28d苯染毒小鼠血清生物标志物的研究

Identification of serum biomarkers change in benzene-exposed mice for 28 days

目的观察28 d苯染毒小鼠血清蛋白的表达变化,寻找其敏感生物标志物。方法选择4~5周龄雄性无特定病原体(specific pathogen free,SPF)级昆明小鼠40只,将其随机分为苯中毒组和对照组,苯中毒组30只,给予苯皮下注射,1次/d,剂量为每千克体质量150 mg(即150 mg/kg),每周5 d,连续4周。对照组10只,按同样方法给予玉米油。染毒期间记录小鼠皮毛外形、精神、饮食状况和体质量变化。实验结束时采集血液,进行血常规检测、血清生化肝功能指标检测、α1-抗胰蛋白酶(α1-AT)、载脂蛋白A1(APOA1)及补体C3的检测;收集小鼠肝脏组织并称重,计算肝脏脏器系数,采用HE染色进行病理分析。结果与对照组比较,苯中毒组实验期间小鼠皮毛外形、精神、饮食状况未见明显异常,染毒4周后,苯中毒组小鼠体质量低于对照组,差异有统计学意义(P <0.05);苯中毒组血红蛋白以及白细胞计数、红细胞计数和血小板计数均低于对照组,差异有统计学意义(P <0.05);苯中毒组的总蛋白、白蛋白、球蛋白、总胆红素、直接胆红素、间接胆红素、谷丙转氨酶、谷草转氨酶、碱性磷酸酶与对照组比较,差异无统计学意义(P> 0.05);肝脏组织病理学检查显示,两组小鼠均有不同程度的肝细胞糖原蓄积;苯中毒组APOA1水平高于对照组,差异有统计学意义(P <0.05)。结论皮下注射是建立苯染毒小鼠模型的简单可行的方法。在苯中毒小鼠模型中,APOA1蛋白表达上调,可能是潜在的苯中毒敏感生物标志物。

Objective To explore the changes of serum protein expression and its sensitive biomarkers in benzene-exposed mice for 28 days. Methods Totally 40 male SPF-grade Kunming mice aged 4 to 5 weeks were randomly divided into benzene poisoning group(n = 30)and control group(n = 10). Mice in benzene poisoning group were given benzene(150 mg/kg)by subcutaneous injection once a day,5 days a week,for 4 weeks. Mice in control group were given corn oil with the same way. During the exposure period,the appearance,mentality,diet and weight changes of mice were recorded. At the end of the experiment,blood was collected for routine blood test and examination of serum biochemical liver function indicators,α1-antitrypsin(α1-AT),apolipoprotein A1(APOA1)and complement C3(C3). Mouse liver organ was taken and weight. The coefficient of liver organs was calculated,and HE staining was done for pathological examination. Results Compared with the control group,the mice in the benzene poisoning group showed no obvious abnormalities in their fur appearance,mentality,and diet during the experiment period. But 4 weeks after exposure,the weight of the mice in the benzene poisoning group was lower than that of the control group,and the difference was statistically significant(P <0.05). The blood test results showed that the white blood cell count,red blood cell count,hemoglobin amount and platelet count of mice in the benzene poisoning group were significantly lower than those of mice in the control group,and the difference was statistically significant(P < 0.05). There was no significant difference in total protein,albumin,globulin,total bilirubin,direct bilirubin,indirect bilirubin,alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase between mice in benzene poisoning group and in control group(P > 0.05). Liver histopathological examination showed that mice in both two groups had somewhat hepatocyte glycogen accumulation. The levels of APOA1 of mice in the benzene poisoning group were higher than those of mice in the control group(P < 0.05). Conclusions Subcutaneous injection is simple and feasible method to induce benzene-poisoning model in mice. In benzene poisoning mice,APOA1 protein expression was up-regulated,and APOA1 may be a potential sensitive biomarker of benzene poisoning.